Background: In December 2016, the Cameroon ministry of Public health in collaboration with WHO updated its HIV guidelines to a test and treat all strategy, expanding antiretroviral therapy (ART) eligibility to all individuals with HIV infection, regardless of CD4+ cell count, and recommending ART be initiated within two weeks of HIV diagnosis and this has been implemented in Cameroon since 2016. Objective: The overall objective of this study was to assess the uptake of universal test and treat strategy and associated challenges. Methods: This was a cross sectional study where participants were randomly selected from 8 communities and 4 Health facilities within Fako Health districts.1501 and 384 participants were randomly selected from the communities and health facilities, respectively .Data was collected using electronic questionnaires and analyzed using SPSS version 25. Chi square test was used to compared proportions between categorical variables while descriptive analysis was used to measure the uptake of Universal test and treat strategy. Results: A total of 1501 respondents were interviewed in the 8 randomly selected communities among which there were 882(58.8%) females and 619(41.1%) males. Among the 384 participants that were sampled from the 4 different health facilities,282(73.4%) and 102(26.6%) were males. With respect to history of HIV test, 1207(85.9%) reported to have ever done an HIV test in their lifetime among which majority (61%) were females and the difference was statistically significant (x2=40.1, p<0.001).Out of the 774 respondents who reported to have visited health facility in the past 12 months, only 517(69.5%) were offer HIV test which was far lower than the expected 100% in the context of universal test and treat strategy. Also, the proportion of females who accepted HIV test was significantly higher than that of the males (66.7% Vs 33.7, P<0.012).With respect to reasons to have taken an HIV test in the past 12 months, majority (62%) indicated voluntary testing, 30.7% reported that it was requested by the health personels. Also, majority (22.8%) of the respondents reported fear of stigmatization as a major reason why they do not want to go in for an HIV test. Major challenges identified in the implementation of test and treat strategy were: Limited number of Psychosocial counsellors (13%), and Poor retention in care (34%). Conclusion: The uptake of the universal test and treat strategy is low and it not implemented across all facilities. Fear remains a major barrier to HIV screening. However, additional research and health promotion advocacy work should be done not only to decrease the fear associated with HIV screening, but also to increase the awareness of the benefits of an early diagnosis, including the effectiveness of the treatment on one’s health and the reduction of transmission to one’s sexual partner. Health workers should scale up the universal test and treat strategy to bring the pandemic under control by 2030.